Size-dependent Mechanisms of Lipid Nanoparticle Induced Endosomal Escape
(2025) KIMM01 20251Department of Immunotechnology
Educational programmes, LTH
- Abstract
- Molecular pathologies can be targeted using lipid nanoparticles carrying therapeutic RNA. Extrahepatic delivery remains a major bottleneck, however, small lipid nanoparticles could allow targeting of deeper tissue layers, low vascularized tumors and the lymph nodes, improving the range of druggable cell types and diseases. Lipid nanoparticle size as a tunable parameter influencing therapeutic efficiency has received little attention, with the mechanism of uptake and endosomal escape of such systems not being fully elucidated. Here we present an initial investigation into how lipid nanoparticle size influences therapeutic efficiency, using confocal microscopic imaging and flow cytometry approaches. We quantified lipid nanoparticle-induced... (More)
- Molecular pathologies can be targeted using lipid nanoparticles carrying therapeutic RNA. Extrahepatic delivery remains a major bottleneck, however, small lipid nanoparticles could allow targeting of deeper tissue layers, low vascularized tumors and the lymph nodes, improving the range of druggable cell types and diseases. Lipid nanoparticle size as a tunable parameter influencing therapeutic efficiency has received little attention, with the mechanism of uptake and endosomal escape of such systems not being fully elucidated. Here we present an initial investigation into how lipid nanoparticle size influences therapeutic efficiency, using confocal microscopic imaging and flow cytometry approaches. We quantified lipid nanoparticle-induced endosomal damages, a necessary step for therapeutic efficiency. At the same time, cellular uptake, protein expression, and gene knockdown were explored to obtain representative insights into the therapeutic efficiency in vitro. The results show that medium (~60 nm) and large (~90-120 nm) lipid nanoparticles outperformed small (~40 nm) ones in protein expression and gene silencing. Small lipid nanoparticles were poorly internalized when loaded with mRNA and in general, caused mostly nonproductive endosomal damages. Despite improved uptake with siRNA as payload, efficiency remained low, suggesting that uptake alone is insufficient without productive endosomal damage and escape. These results pave the way to a shift in the perception of small lipid nanoparticles, focusing on alterations to the nanoparticle composition to improve endosomal escape and thus therapeutic efficiency, improving its clinical application. (Less)
- Popular Abstract
- Size Matters: Investigating lipid nanoparticle size for improved treatment efficiency
Imagine a world where diseases like cancers and genetic defects could be easily treated. Whilst we are still far from this utopian futuristic scenario, lipid nanoparticles have emerged as elegant and effective drug carriers for treatment delivery. Their cargo can vary from small molecular drugs, through “genetic” medicines like mRNA and siRNA up to whole protein complexes such as CRISPR-Cas9 to correct genetic mistakes. In addition, their lipid composition and other properties such as size are tunable which allows for greater freedom and targeting of different tissues within the organism.
Currently, lipid nanoparticles predominantly target the... (More) - Size Matters: Investigating lipid nanoparticle size for improved treatment efficiency
Imagine a world where diseases like cancers and genetic defects could be easily treated. Whilst we are still far from this utopian futuristic scenario, lipid nanoparticles have emerged as elegant and effective drug carriers for treatment delivery. Their cargo can vary from small molecular drugs, through “genetic” medicines like mRNA and siRNA up to whole protein complexes such as CRISPR-Cas9 to correct genetic mistakes. In addition, their lipid composition and other properties such as size are tunable which allows for greater freedom and targeting of different tissues within the organism.
Currently, lipid nanoparticles predominantly target the liver and liver associated diseases, however smaller lipid nanoparticles are expected to have the advantage of squeezing through narrow vessels and into dense tissues targeting in other ways hard to reach cells. That is why by altering the size of lipid nanoparticles, all bodily tissues should eventually become druggable.
We have set out to investigate the therapeutic efficiency of different sized lipid nanoparticles. For the treatment to be efficient, the drug carrier must first pass the cell membrane and subsequently escape from an encapsulated vesicle called the endosome into the cytosol. Hence, cell uptake and endosomal escape, coupled with protein expression (for mRNA cargo) or gene silencing (for siRNA cargo) were our measured parameters. Our results surprised us, as the smallest particles (about 40 nanometers) didn’t show good results. They were taken up poorly, especially when transporting mRNA, and often caused only few unproductive damages. Uptake was improved when siRNA was transported, however this change did not improve the gene silencing ability. Medium (60 nm) and large (90–120 nm) nanoparticles performed better, successfully delivering their therapeutic payloads and triggering changes inside cells, partly due to their larger transportation volume and partly due to inducing more successful damages.
These findings highlight that how particles are built up in addition to size can make or break a treatment. This insight could help improve the design of future RNA-based drugs for hard-to-reach areas like tumors or lymph nodes, opening new doors in personalized medicine and cancer therapy. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9195130
- author
- Hains, Patrik LU
- supervisor
- organization
- course
- KIMM01 20251
- year
- 2025
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Biotechnology, RNA therapeutics, Endosomal escape, Lipid nanoparticles, Cancer Therapy, Gene therapy
- language
- English
- id
- 9195130
- date added to LUP
- 2025-06-19 15:26:21
- date last changed
- 2025-06-19 15:26:21
@misc{9195130,
abstract = {{Molecular pathologies can be targeted using lipid nanoparticles carrying therapeutic RNA. Extrahepatic delivery remains a major bottleneck, however, small lipid nanoparticles could allow targeting of deeper tissue layers, low vascularized tumors and the lymph nodes, improving the range of druggable cell types and diseases. Lipid nanoparticle size as a tunable parameter influencing therapeutic efficiency has received little attention, with the mechanism of uptake and endosomal escape of such systems not being fully elucidated. Here we present an initial investigation into how lipid nanoparticle size influences therapeutic efficiency, using confocal microscopic imaging and flow cytometry approaches. We quantified lipid nanoparticle-induced endosomal damages, a necessary step for therapeutic efficiency. At the same time, cellular uptake, protein expression, and gene knockdown were explored to obtain representative insights into the therapeutic efficiency in vitro. The results show that medium (~60 nm) and large (~90-120 nm) lipid nanoparticles outperformed small (~40 nm) ones in protein expression and gene silencing. Small lipid nanoparticles were poorly internalized when loaded with mRNA and in general, caused mostly nonproductive endosomal damages. Despite improved uptake with siRNA as payload, efficiency remained low, suggesting that uptake alone is insufficient without productive endosomal damage and escape. These results pave the way to a shift in the perception of small lipid nanoparticles, focusing on alterations to the nanoparticle composition to improve endosomal escape and thus therapeutic efficiency, improving its clinical application.}},
author = {{Hains, Patrik}},
language = {{eng}},
note = {{Student Paper}},
title = {{Size-dependent Mechanisms of Lipid Nanoparticle Induced Endosomal Escape}},
year = {{2025}},
}